Uropathogenic Escherichia coli (UPEC) causes uncomplicated urinary tract infection (UTI) depicts a prevalent and potentially uncompromising infectious disease. In this analysis, we explained the functions of an immunoproteomics concept to vaccine development that has been successfully employed to recognize vaccine targets in other pathogenic bacteria. Pyelonephritis strains E. coli CFT073 are used for outer membrane isolation mimics urinary tract environment in which iron limitation, osmotic stress, human urine, and exposure to uroepithelial cells are included. During experiments of UTI, the antigens that induce the humoral immune response is to identified, two-dimensional gel electrophoresis are employed for the isolation of outer membrane protein and probed using pooled antisera from 20 CBA/J mice chronically infected with E. coli CFT073. 23 total outer membrane antigens, in which a unique iron compound receptor is included, are reacted with antisera and were identified by mass spectrometry. These antigens comprises of proteins with known functions in UPEC pathogenesis such as, ChuA, IroN, IreA, Iha, IutA, and FliC. These all information and data elaborate that these factors are associated with virulence during UTI are directed by antibody response. We also represents that the genes encoding ChuA, IroN, hypothetical protein c2482, and IutA are significantly more prevalent among UPEC strains than among fecal-commensal E. coli isolates. Therefore we concluded that, the outer membrane antigens are identified in this study are conserved, could be reflective part for the UTI vaccine generated to induce protective immunity against UPEC infections.